Hinoda Y, Nakagawa N, Nakamura H, Makiguchi Y, Itoh F, Adachi M, Yabana T, Imai K, Yachi A
Department of Internal Medicine (Section 1), Sapporo Medical College, Japan.
Immunol Lett. 1993 Feb;35(2):163-8. doi: 10.1016/0165-2478(93)90086-h.
This study aims at clarifying whether the humoral immune response to the tandem repeat domain of MUC1 can be induced or not in vivo. The expression of MUC1 mRNA in the colon was revealed by Northern blot analysis, and cDNA cloning of an extracellular tandemly repeated domain of MUC1 was then performed to prepare the recombinant MUC1 protein. A cDNA clone coding for ten repeat domains was ligated into an expression vector in prokaryotes, resulting in a recombinant protein which could react with the MAb MUSE11 against an adenocarcinoma-associated antigen whose epitope has been shown to be localized in the tandem repeat domain on MUC1. The reactivity of sera from patients with ulcerative colitis with the recombinant protein was evaluated by SDS-PAGE and Western blot analysis to detect the antibodies against this tandem repeat domain. Five out of 19 serum samples tested positive, and these reactions were totally inhibited by MAb MUSE11, suggesting that the epitope recognized by these antibodies in sera is almost identical to that recognized by MAb MUSE11. The data represent the first demonstration of antibody production against a peptide epitope of the tandem repeat domain of MUC1.
本研究旨在阐明在体内是否能够诱导针对MUC1串联重复结构域的体液免疫反应。通过Northern印迹分析揭示了结肠中MUC1 mRNA的表达，随后进行MUC1细胞外串联重复结构域的cDNA克隆，以制备重组MUC1蛋白。将编码十个重复结构域的cDNA克隆连接到原核表达载体中，产生一种重组蛋白，该蛋白可与抗腺癌相关抗原的单克隆抗体MUSE11发生反应，其表位已显示定位于MUC1的串联重复结构域上。通过SDS-PAGE和Western印迹分析评估溃疡性结肠炎患者血清与重组蛋白的反应性，以检测针对该串联重复结构域的抗体。19份检测的血清样本中有5份呈阳性，并且这些反应被单克隆抗体MUSE11完全抑制，这表明血清中这些抗体识别的表位与单克隆抗体MUSE11识别的表位几乎相同。这些数据首次证明了针对MUC1串联重复结构域的肽表位产生了抗体。
